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1.
Nature ; 594(7862): 271-276, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33910229

RESUMO

Vascular malformations are thought to be monogenic disorders that result in dysregulated growth of blood vessels. In the brain, cerebral cavernous malformations (CCMs) arise owing to inactivation of the endothelial CCM protein complex, which is required to dampen the activity of the kinase MEKK31-4. Environmental factors can explain differences in the natural history of CCMs between individuals5, but why single CCMs often exhibit sudden, rapid growth, culminating in strokes or seizures, is unknown. Here we show that growth of CCMs requires increased signalling through the phosphatidylinositol-3-kinase (PI3K)-mTOR pathway as well as loss of function of the CCM complex. We identify somatic gain-of-function mutations in PIK3CA and loss-of-function mutations in the CCM complex in the same cells in a majority of human CCMs. Using mouse models, we show that growth of CCMs requires both PI3K gain of function and CCM loss of function in endothelial cells, and that both CCM loss of function and increased expression of the transcription factor KLF4 (a downstream effector of MEKK3) augment mTOR signalling in endothelial cells. Consistent with these findings, the mTORC1 inhibitor rapamycin effectively blocks the formation of CCMs in mouse models. We establish a three-hit mechanism analogous to cancer, in which aggressive vascular malformations arise through the loss of vascular 'suppressor genes' that constrain vessel growth and gain of a vascular 'oncogene' that stimulates excess vessel growth. These findings suggest that aggressive CCMs could be treated using clinically approved mTORC1 inhibitors.


Assuntos
Classe I de Fosfatidilinositol 3-Quinases/genética , Hemangioma Cavernoso do Sistema Nervoso Central/genética , Hemangioma Cavernoso do Sistema Nervoso Central/patologia , Mutação , Neoplasias/genética , Animais , Animais Recém-Nascidos , Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Mutação com Ganho de Função , Hemangioma Cavernoso do Sistema Nervoso Central/irrigação sanguínea , Hemangioma Cavernoso do Sistema Nervoso Central/metabolismo , Humanos , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/metabolismo , Mutação com Perda de Função , MAP Quinase Quinase Quinase 3/metabolismo , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina/antagonistas & inibidores , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Camundongos , Neoplasias/irrigação sanguínea , Neoplasias/patologia , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/metabolismo
2.
EMBO J ; 41(9): e111189, 2022 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-35437788

RESUMO

A functional blood-brain barrier relies on a tightly controlled interplay between endothelial cells, pericytes, and astrocytes, which together form the neurovascular unit. Recent work by Lee et al (2022) discovers endothelial cell-derived lactate as a crucial metabolic fuel for brain pericytes, revealing a new way of CNS vascular communication that links nutrient metabolism to blood-brain barrier function.


Assuntos
Barreira Hematoencefálica , Células Endoteliais , Astrócitos , Encéfalo , Ecossistema , Pericitos/metabolismo
3.
Nucleic Acids Res ; 52(13): e56, 2024 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-38850155

RESUMO

Methods for modifying gene function at high spatiotemporal resolution in mice have revolutionized biomedical research, with Cre-loxP being the most widely used technology. However, the Cre-loxP technology has several drawbacks, including weak activity, leakiness, toxicity, and low reliability of existing Cre-reporters. This is mainly because different genes flanked by loxP sites (floxed) vary widely in their sensitivity to Cre-mediated recombination. Here, we report the generation, validation, and utility of iSuRe-HadCre, a new dual Cre-reporter and deleter mouse line that avoids these drawbacks. iSuRe-HadCre achieves this through a novel inducible dual-recombinase genetic cascade that ensures that cells expressing a fluorescent reporter had only transient Cre activity, that is nonetheless sufficient to effectively delete floxed genes. iSuRe-HadCre worked reliably in all cell types and for the 13 floxed genes tested. This new tool will enable the precise, efficient, and trustworthy analysis of gene function in entire mouse tissues or in single cells.


Assuntos
Integrases , Animais , Integrases/genética , Integrases/metabolismo , Camundongos , Genes Reporter , Recombinação Genética
4.
Parasitology ; 150(2): 172-183, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36444644

RESUMO

Helminth species of Neotropical bats are poorly known. In Mexico, few studies have been conducted on helminths of bats, especially in regions such as the Yucatan Peninsula where Chiroptera is the mammalian order with the greatest number of species. In this study, we characterized morphologically and molecularly the helminth species of bats and explored their infection levels and parasite­host interactions in the Yucatan Peninsula, Mexico. One hundred and sixty-three bats (representing 21 species) were captured between 2017 and 2022 in 15 sites throughout the Yucatan Peninsula. Conventional morphological techniques and molecular tools were used with the 28S gene to identify the collected helminths. Host­parasite network analyses were carried out to explore interactions by focusing on the level of host species. Helminths were found in 44 (26.9%) bats of 12 species. Twenty helminth taxa were recorded (7 trematodes, 3 cestodes and 10 nematodes), including 4 new host records for the Americas. Prevalence and mean intensity of infection values ranged from 7.1 to 100% and from 1 to 56, respectively. Molecular analyses confirmed the identity of some helminths at species and genus levels; however, some sequences did not correspond to any of the species available on GenBank. The parasite­host network suggests that most of the helminths recorded in bats were host-specific. The highest helminth richness was found in insectivorous bats. This study increases our knowledge of helminths parasitizing Neotropical bats, adding new records and nucleotide sequences.


Assuntos
Quirópteros , Helmintíase Animal , Helmintos , Nematoides , Parasitos , Animais , Quirópteros/parasitologia , México/epidemiologia , Helmintos/genética , Interações Hospedeiro-Parasita , Helmintíase Animal/epidemiologia , Helmintíase Animal/parasitologia
5.
Scand J Immunol ; 95(4): e13143, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35067952

RESUMO

INTRODUCTION: For many patients with primary immune deficiency (PID), stem-cell transplantation (SCT) may be life-saving. OBJECTIVE: To review our experience of 11 years transplanting children with PID in Mexico. METHODS: Chart review of patients who underwent SCT from 2008 to 2018, to describe their diagnoses, time to transplant, conditioning regime, survival rate and outcomes. All patients received post-transplant cyclophosphamide as graft-versus-host-disease (GVHD) prophylaxis. RESULTS: 19 patients with combined, phagocytic or syndromic PID from 5 states. Twelve of them were male (58%) and 14 survive (79%). Mean age at HSCT was 41.9 months; mean time from diagnosis was 31.2 months. Seven grafts were umbilical cord and 12 haploidentical. The conditioning regime was myeloablative, with five primary graft failures. Two patients had partial and 10 full chimerism. Five patients died within 2 months after transplant. Immune reconstitution was complete in 11 of 19 patients. We found a prevalence of 21% GVHD. DISCUSSION: We describe 19 patients from Mexico with 8 PID diagnoses who underwent allogenic HSCT over a period of 11 years. Survival rate and other outcomes compare well with industrialized countries. We recommend the use of post-transplant cyclophosphamide to prevent GVHD in scenarios of resource scarcity and a lack of HLA-identical donors.


Assuntos
Doença Enxerto-Hospedeiro , Transplante de Células-Tronco Hematopoéticas , Doenças da Imunodeficiência Primária , Criança , Ciclofosfamida/uso terapêutico , Doença Enxerto-Hospedeiro/prevenção & controle , Humanos , Masculino , México , Doenças da Imunodeficiência Primária/terapia , Estudos Retrospectivos , Condicionamento Pré-Transplante
6.
Int J Trop Insect Sci ; 42(2): 2007-2012, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34745312

RESUMO

After the tropical storm Cristobal, we performed special adult entomological collections in the peri-domicile of 35 houses from 25 neighborhoods of Mérida, Yucatan, Mexico in response to complaints from the community about an increased nuisance due to an abundance of mosquitoes. A total of 1,275 specimens from four genera and 13 species were collected: Aedes taeniorhynchus (92%), Culex quinquefasciatus (72%), Aedes aegypti (72%), Psorophora mexicana (36%), Psorophora cyanescens (32%), Aedes scapularis (24%), Culex nigripalpus (24%), Aedes albopictus (8%), Psorophora ferox (4%), Haemagogus equinus (4%), Aedes trivittatus (4%), Culex coronator (4%), Culex iolambdis (4%). From these collections, the increased mosquito nuisance was mainly the result of invasive species such as Aedes taeniorhynchus and Psorophora. City wide, vehicle mounted ULV spraying was performed by the MoH and the municipality of Merida to control adult mosquito populations. We report Culex iolambdis for the first time in Merida and Psorophora mexicana for the state of Yucatan.

7.
Development ; 145(13)2018 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-29853619

RESUMO

Formation and homeostasis of the vascular system requires several coordinated cellular functions, but their precise interplay during development and their relative importance for vascular pathologies remain poorly understood. Here, we investigated the endothelial functions regulated by Cdc42 and their in vivo relevance during angiogenic sprouting and vascular morphogenesis in the postnatal mouse retina. We found that Cdc42 is required for endothelial tip cell selection, directed cell migration and filopodia formation, but dispensable for cell proliferation or apoptosis. Although the loss of Cdc42 seems generally compatible with apical-basal polarization and lumen formation in retinal blood vessels, it leads to defective endothelial axial polarization and to the formation of severe vascular malformations in capillaries and veins. Tracking of Cdc42-depleted endothelial cells in mosaic retinas suggests that these capillary-venous malformations arise as a consequence of defective cell migration, when endothelial cells that proliferate at normal rates are unable to re-distribute within the vascular network.


Assuntos
Capilares/anormalidades , Movimento Celular , Células Endoteliais/metabolismo , Veia Retiniana/anormalidades , Malformações Vasculares/embriologia , Proteína cdc42 de Ligação ao GTP/deficiência , Animais , Capilares/embriologia , Polaridade Celular/genética , Células Endoteliais/patologia , Camundongos , Camundongos Knockout , Pseudópodes/genética , Pseudópodes/metabolismo , Veia Retiniana/embriologia , Malformações Vasculares/genética , Malformações Vasculares/patologia
8.
Circ Res ; 124(8): 1240-1252, 2019 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-30732528

RESUMO

RATIONALE: Aberrant formation of blood vessels precedes a broad spectrum of vascular complications; however, the cellular and molecular events governing vascular malformations are not yet fully understood. OBJECTIVE: Here, we investigated the role of CDC42 (cell division cycle 42) during vascular morphogenesis and its relative importance for the development of cerebrovascular malformations. METHODS AND RESULTS: To avoid secondary systemic effects often associated with embryonic gene deletion, we generated an endothelial-specific and inducible knockout approach to study postnatal vascularization of the mouse brain. Postnatal endothelial-specific deletion of Cdc42 elicits cerebrovascular malformations reminiscent of cerebral cavernous malformations (CCMs). At the cellular level, loss of CDC42 function in brain endothelial cells (ECs) impairs their sprouting, branching morphogenesis, axial polarity, and normal dispersion within the brain tissue. Disruption of CDC42 does not alter EC proliferation, but malformations occur where EC proliferation is the most pronounced during brain development-the postnatal cerebellum-indicating that a high, naturally occurring EC proliferation provides a permissive state for the appearance of these malformations. Mechanistically, CDC42 depletion in ECs elicited increased MEKK3 (mitogen-activated protein kinase kinase kinase 3)-MEK5 (mitogen-activated protein kinase kinase 5)-ERK5 (extracellular signal-regulated kinase 5) signaling and consequent detrimental overexpression of KLF (Kruppel-like factor) 2 and KLF4, recapitulating the hallmark mechanism for CCM pathogenesis. Through genetic approaches, we demonstrate that the coinactivation of Klf4 reduces the severity of vascular malformations in Cdc42 mutant mice. Moreover, we show that CDC42 interacts with CCMs and that CCM3 promotes CDC42 activity in ECs. CONCLUSIONS: We show that endothelial-specific deletion of Cdc42 elicits CCM-like cerebrovascular malformations and that CDC42 is engaged in the CCM signaling network to restrain the MEKK3-MEK5-ERK5-KLF2/4 pathway.


Assuntos
Vasos Sanguíneos/anormalidades , Proliferação de Células , Células Endoteliais/fisiologia , Deleção de Genes , Hemangioma Cavernoso do Sistema Nervoso Central/etiologia , Proteína cdc42 de Ligação ao GTP/genética , Animais , Animais Recém-Nascidos , Proteínas Reguladoras de Apoptose/genética , Encéfalo/irrigação sanguínea , Ciclo Celular/fisiologia , Proteína KRIT1/genética , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/metabolismo , MAP Quinase Quinase 5/metabolismo , MAP Quinase Quinase Quinase 3/metabolismo , Camundongos , Proteínas dos Microfilamentos/genética , Proteína cdc42 de Ligação ao GTP/metabolismo
9.
Resour Conserv Recycl ; 164: 105152, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32921916

RESUMO

The Covid-19 pandemic will leave a lasting impact on nearly every aspect of life in society and has also raised concerns on the contamination risks associated with waste management. This study presents a review on the main recommendations related to the management of municipal solid wastes during the pandemic. The recommendations were classified according to the target audience, and their applicability to the Brazilian reality is discussed. Results show that most analyzed recommendations are related to hygiene routines, use of personal protective equipment, and proper segregation, packing and final destination of potentially contaminated wastes. Brazilian organizations show a special concern about the exposure risks of waste pickers, advising for the temporary suspension of manual waste collection and sorting, hygiene protocols, social distancing and quarantine of recyclable materials. Overall, the recommendations are highly dependent on awareness and engagement of citizens and on planning and support of municipalities, which must invest in information campaigns and provide alternatives for the infectious waste produced in households. Furthermore, this study points out that the recommendations were issued in an emergency scenario, but as the pandemic is expected to last for an extended period of time, they must be revised as local contexts change, seeking to maintain and extend citizens engagement in selective collection and even in actions towards waste reduction.

10.
An Acad Bras Cienc ; 92(1): e20180193, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32267302

RESUMO

The isotopic and geochemical evolution of tourmaline constrain the processes of paleo-oceanic lithosphere in ophiolites. The Brasiliano Orogen is a major structure of South America and requires characterization for the understanding of Gondwana supercontinent evolution. We made a pioneering investigation of tourmaline from a tourmalinite in the Ibaré ophiolite by integrating field work with chemical analyses of tourmaline by electron microprobe (EPMA) and δ11B determinations via laser ablation inductively coupled plasma mass spectrometer (LA-ICP-MS). Remarkably massive tourmalinite (>90 vol.% tourmaline, some chlorite) enclosed in serpentinite has homogeneous dravite in chemical and isotopic composition (δ11B = +3.5 to +5.2‰). These results indicate a geotectonic environment in the altered oceanic crust for the origin of the tourmalinite. This first δ11B characterization of tourmaline from tourmalinite sets limits to the evolution of the Neoproterozoic to Cambrian Brasiliano Orogen and Gondwana evolution.

12.
Vet Parasitol Reg Stud Reports ; 53: 101077, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39025538

RESUMO

The monthly fluctuation of adult Ixodes keiransi in dogs from two cattle farms in southeastern Mexico was determined. In ranch 1 (R1), 6-7 mixed breed dogs from six months to five years of age; while in ranch 2 (R2), 7-8 mixed breed dogs from one to four years of age, were followed. All dogs were monthly inspected for tick infestation through one year. The abundance of dogs infested with ticks was estimated for each month. Tick specimens were taxonomically identified as I. keiransi using morphological features. In R1 the dogs were inspected 60 times and in R2 90 times. The overall proportion of dogs infested by ticks through the year was 75% (60/80) in R1 and 67% (67/90) in R2. The higher abundance of adult I. keiransi parasitizing dogs was from July to December with maximum peaks from September to November. In April, May and June, no ticks were observed on dogs from both ranches. Tick abundance fluctuation showed a concordant pattern with rainfall but with a delay because de maximum peak of I. keiransi abundance was almost two months after the greatest rainfall peak. It is concluded that, in some conditions, the adult I. keiransi in southeastern Mexico presents high-infested rates on dogs and a maximum abundance from September to November. The I. keiransi abundance fluctuation showed a concordant pattern with rainfall.


Assuntos
Doenças do Cão , Ixodes , Estações do Ano , Infestações por Carrapato , Animais , Cães , México/epidemiologia , Infestações por Carrapato/veterinária , Infestações por Carrapato/parasitologia , Infestações por Carrapato/epidemiologia , Doenças do Cão/parasitologia , Doenças do Cão/epidemiologia , Ixodes/fisiologia , Ixodes/classificação , Ixodes/parasitologia , Feminino , Masculino , Bovinos/parasitologia
13.
J Hazard Mater ; 469: 133955, 2024 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-38457976

RESUMO

The complexity around the dynamic markets for new psychoactive substances (NPS) forces researchers to develop and apply innovative analytical strategies to detect and identify them in influent urban wastewater. In this work a comprehensive suspect screening workflow following liquid chromatography - high resolution mass spectrometry analysis was established utilising the open-source InSpectra data processing platform and the HighResNPS library. In total, 278 urban influent wastewater samples from 47 sites in 16 countries were collected to investigate the presence of NPS and other drugs of abuse. A total of 50 compounds were detected in samples from at least one site. Most compounds found were prescription drugs such as gabapentin (detection frequency 79%), codeine (40%) and pregabalin (15%). However, cocaine was the most found illicit drug (83%), in all countries where samples were collected apart from the Republic of Korea and China. Eight NPS were also identified with this protocol: 3-methylmethcathinone 11%), eutylone (6%), etizolam (2%), 3-chloromethcathinone (4%), mitragynine (6%), phenibut (2%), 25I-NBOH (2%) and trimethoxyamphetamine (2%). The latter three have not previously been reported in municipal wastewater samples. The workflow employed allowed the prioritisation of features to be further investigated, reducing processing time and gaining in confidence in their identification.


Assuntos
Drogas Ilícitas , Poluentes Químicos da Água , Águas Residuárias , Fluxo de Trabalho , Psicotrópicos , China , Poluentes Químicos da Água/análise
14.
Acta Trop ; 251: 107113, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38157924

RESUMO

Bats are one of the groups of mammals with the highest number of associated Trypanosoma taxa. There are 50 Trypanosoma species and genotypes infecting more than 75 species of bats across five continents. However, in Mexico, the inventory of species of the genus Trypanosoma associated with bats is limited to only two species (Trypanosoma vespertilionis and Trypanosoma cruzi) even though 140 species of bats inhabit this country. Specifically, 91 bat species have been recorded in the state of Veracruz, but records of trypanosomatids associated with this mammalian group are absent. Due to the complex Trypanosoma-bat relationship, the high diversity of bat species in Veracruz, as well as the lack of records of trypanosomatids associated with bats for this state, the aim of this work was to analyze the diversity of species of the genus Trypanosoma and their presence from a bat community in the central area of the state of Veracruz, Mexico. During the period of January to August 2022 in the Tequecholapa Environmental Management Unit where bats were collected using mist nets and blood samples were obtained from their thumbs. We extracted genetic material and amplified a fragment of 800 bp of the 18S ribosomal gene of the genus Trypanosoma by conventional PCR. The positive amplicons were sequenced, and phylogenetic reconstruction was performed to identify the parasite species. A total of 285 bats (149♀, 136♂) belonging to 13 species from 10 genera and a single family (Phyllostomidae) were collected. Twenty-three specimens from six species tested positive for the presence of Trypanosoma dionisii, Trypanosoma sp. Neobat 4, and a potential novelty species provisionally named as Trypanosoma sp. Neobat 6. The results of the present work increase the number of species of the genus Trypanosoma infecting bats in Mexico and in the Neotropical region.


Assuntos
Quirópteros , Trypanosoma cruzi , Trypanosoma , Animais , Quirópteros/parasitologia , Filogenia , México , Trypanosoma/genética , Trypanosoma cruzi/genética , Sequência de Bases
15.
Acta Trop ; 249: 107072, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38008370

RESUMO

This study aims to describe the natural Leptospira occurrence in small mammals from Yucatan, Mexico, and to explore the relation between the characteristics of the capture sites and the Leptospira occurrence. Bats and rodents were captured in five sites of Yucatan state, and from them, a kidney fragment was collected that was used in the genomic DNA extraction. Leptospira DNA was identified by PCR targeting the 16S-rRNA and LipL32 genes. Additionally, a bioinformatic analysis was carried out to know the Leptospira species and was corroborated with a phylogenetic tree. The assemblage of small mammals was compound of 82 (51.2 %) bats and 78 (48.8 %) rodents. A global frequency (bats plus rodents) of Leptospira occurrence of 21.2 % (34/160) was observed; in bats, it was 21.9 % (18/82), and in rodents, 20.5 % (16/78). The phylogenetic trees based on LipL32 gene showed that the recovered sequences most closely resemble the species L. borgpetersenii and L. noguchii. The ordination of the capture sites with tropical deciduous forests as original vegetation is more related to the abundance of Leptospira-infected rodents. The ordination of the capture sites with tropical sub-deciduous forests as original vegetation is more related to the diversity of Leptospira-infected bat species. The canonical ordering of the capture sites is by the original vegetation type and the diversity and abundance of Leptospira-infected bat and rodent species.


Assuntos
Quirópteros , Leptospira , Leptospirose , Animais , Leptospira/genética , Leptospirose/epidemiologia , Leptospirose/veterinária , México/epidemiologia , Roedores , Filogenia , DNA Bacteriano/genética
16.
Braz J Microbiol ; 54(1): 151-158, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36574206

RESUMO

BACKGROUND: Chlamydia trachomatis (CT) infection is the most prevalent sexually transmitted bacterial disease worldwide whose greatest impact is on the female reproductive system. The objective was to assess the prevalence of Chlamydia infection in a large population of female patients from a private health service in São Paulo (Brazil), identifying the main age groups affected by the agent and the evolution of the prevalence. METHODS: A cross-sectional study was conducted based on the results of all molecular biology tests. The tests were carried out between January 2005 and December 2015. The positivity of test results was determined by year and age group. RESULTS: A total of 85,292 patients who performed 103,576 tests for Chlamydia were considered eligible for the statistical data. The overall prevalence of C. trachomatis infection in the study population was 2.2% (95% CI: 2.07-2.25). A higher prevalence of infection was observed in the ≤ 25 years and ≤ 30 years age groups, with rates of 6.0% (95% CI: 5.59-6.35) and 4.4% (95% CI: 4.08-4.50), respectively. There was a significant increase in the positivity of the exams over time, especially in the ≤ 35 years age group. The prevalence at 26-30 years was 3.1% (95% CI: 2.82-3.30); 31-35 years 1.7% (95% CI: 1.50-1.82); 36-40 years 1.0% (95% CI: 0.86-1.16); 41-60 years 0.6% (95% CI: 0.50-0.70) and the prevalence at ≥ 61 years was 0.4% (95% CI: 0.11-0.75). CONCLUSION: The screening of asymptomatic young women would have the potential to reduce infection, transmission, and sequelae of infection by this agent.


Assuntos
Infecções por Chlamydia , Chlamydia trachomatis , Humanos , Feminino , Prevalência , Brasil/epidemiologia , Estudos Transversais , Infecções por Chlamydia/diagnóstico , Fatores de Risco
17.
Front Microbiol ; 14: 1027380, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36819063

RESUMO

Little is known about the gene expression program during the transition from lysogenic to lytic cycles of temperate bacteriophages in Pseudomonas aeruginosa. To investigate this issue, we developed a thermo-sensitive repressor mutant in a lysogen and analyzed the phage transcriptional program by strand-specific RNA-Seq before and after thermo-induction. As expected, the repressor gene located on the phage DNA forward strand is transcribed in the lysogen at the permissive temperature of 30°C. Upstream the repressor gene, we noticed the presence of two overlapped ORFs apparently in the same transcript. One ORF is a gene that encodes a protein of 7.9 kDa mediating the exclusion of various super-infecting phages. The other ORF, placed in an alternate reading frame with a possible AUG initiation codon at 25 nucleotide downstream of the AUG of the first gene, is expected to encode a 20.7 kDa polypeptide of yet an unknown function. Upon lifting repression at 40°C, the transcription of an operon which is involved in the lytic cycle is started from a promoter on the reverse phage DNA strand. The first gene in the operon is a homolog of the antirepresor ner, a common gene in the lysis-lysogeny regulation region of other phages. Interestingly, the next gene after ner is gene 10 that on the reverse strand overlaps the overlapped gene olg1 on the forward strand. Curiously, gene 10 expression also shows superinfection exclusion. Strand-specific RNA-Seq also has uncovered the transcription succession of gene modules expressed during the phage lytic stage. The conservation of overlapped genes with similar functions may be evolutionarily selected.

18.
Zoonoses Public Health ; 70(7): 594-603, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37391871

RESUMO

Rickettsia parkeri belongs to the spotted fever group (SFG) of the Rickettsia genus. This bacterium causes mild rickettsiosis in humans and is mainly transmitted by Amblyomma ticks. Its medical importance is emerging in the Americas, including Mexico. Synanthropic rodents and domiciled dogs participate as accidental hosts in epidemiological cycles of Rickettsia of the SFG. The aim is to report the presence of R. parkeri in synanthropic rodents and domiciled dogs from a rural community of Yucatán, Mexico. Rodents were captured, and plasma samples were taken from dogs in 48 households from Ucú, Yucatán, Mexico. A spleen sample (rodents) and plasma (dogs) were used in the propagation of Rickettsia on Vero cells. These infected cells were used in the extraction of genomic DNA. Rickettsia DNA was identified using a semi-nested PCR (snPCR); some products were sent for sequencing. The recovered sequences were analysed with bioinformatics programs, and a phylogenetic tree was built to determine the Rickettsia species. One hundred animals were sampled: 36 synanthropic rodents and 64 dogs. The snPCR evidenced the presence of Rickettsia DNA in 10 rodents (10/36, 27.8%) and 18 dogs (18/64, 28.1%), which represents a global frequency of 28% (28/100) in this study. The bioinformatics analysis yielded homology to R. parkeri and was demonstrated in the phylogenetic tree. The first evidence of the presence of R. parkeri in synanthropic rodents (Mus musculus) from Mexico is presented; likewise, the participation of domestic dogs in the transmission cycle of this bacterium with potential importance in public health is confirmed.

19.
Vet Parasitol Reg Stud Reports ; 44: 100911, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37652628

RESUMO

Trypanosoma cruzi, the causal agent of American trypanosomiasis, and Leishmania spp., the causal agents of Leishmaniasis, are prevalent in more than 20 American countries, including Mexico. Dogs have been reported as incidental hosts for both parasites and may be helpful as transmission sentinels. We surveyed the dog population in a rural locality of the Merida municipality in Yucatan, Mexico, to evaluate the seroreactivity against T. cruzi and Leishmania spp. using two antigens, parasite homogenate (H) and iron superoxide dismutase extract (FeSODe), with two serological techniques (ELISA and Western Blot). Our study found that 3.33% of the tested dogs were seroreactive to T. cruzi using ELISA-H, and 29.5% were seroreactive to FeSODe antigen, with a 94.4% consistency between the two tests. Similarly, for L. mexicana, 1.6% were seroreactive using ELISA-H, and 9.8% were seroreactive using ELISA-FeSODe, with an 83.3% consistency between tests. For L. braziliensis, no dogs were seroreactive using ELISA-H, but 16.4% were seroreactive using ELISA-FeSODe, with a 90% consistency between tests. Finally, for L. infantum, 4.9% were seropositive using ELISA-H, and 6.6% were seropositive using ELISA-FeSODe, with a 75% consistency between tests. These results show noticeable evidence of exposure of dogs to trypanosomatid parasites and highlight the potential disease risk for the people and their companion animals in the region.


Assuntos
Doença de Chagas , Leishmania , Parasitos , Trypanosoma cruzi , Animais , México/epidemiologia , Doença de Chagas/epidemiologia , Doença de Chagas/veterinária
20.
Zootaxa ; 5357(2): 205-240, 2023 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-38220646

RESUMO

In this survey, we inventoried the helminths of heteromyid and cricetid rodents captured in the Yucatan Peninsula from 2017 to 2019. Helminths were identified using morphological techniques (clearing, staining, and scanning electron microscopy). Also, the 28S rRNA gene of individuals from several helminth taxa was successfully amplified and sequenced. To confirm the identification at the generic level, and in some cases at the specific level, and the genealogical relationships of the parasites, phylogenetic analyses were performed with the new 28S sequences. We identified 22 species of helminths including three trematodes (Brachylaimidae, Dicrocoeliidae, and Microphallidae), five cestodes (Davaineidae, Hymenolepididae, and Taeniidae), and 14 nematodes (Trichuridae, Ancylostomatidae, Ornithostrongylidae, Heligmonellidae, and Oxyuridae) from Heteromys gaumeri (Heteromyidae), Ototylomys phyllotis, Oligoryzomys fulvescens, Peromyscus yucatanicus, Sigmodon toltecus, and Reithrodontomys gracilis (Cricetidae). The overall frequency of infection in small rodents was 84.1% (143/170); all specimens of H. gaumeri, S. toltecus and Ol. fulvescens were infected with helminths. In total, we provided 46 new sequences of the 28S gene from 17 species of helminths. Seven species are likely undescribed species, six are reported for the first time in rodents from Mexico, and 12 are new host records in the Americas. Before this study, 87 taxa of helminths had been reported from 35 cricetid and 12 heteromyid species in 21 Mexican states. Our findings increase to 93 the helminth taxa in these rodents, and to 36 the cricetid species parasitized by helminths. This large scale-survey is the first to use an integrative approach to inventory the helminths of wild small rodents in Mexico.


Assuntos
Helmintos , Roedores , Animais , Roedores/parasitologia , Arvicolinae/genética , Arvicolinae/parasitologia , México , Filogenia , Helmintos/genética , Sigmodontinae , Peromyscus
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